CN109486881A - A kind of fermentation medium and zymotechnique of kasugarnycin - Google Patents

A kind of fermentation medium and zymotechnique of kasugarnycin Download PDF

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Publication number
CN109486881A
CN109486881A CN201811608846.8A CN201811608846A CN109486881A CN 109486881 A CN109486881 A CN 109486881A CN 201811608846 A CN201811608846 A CN 201811608846A CN 109486881 A CN109486881 A CN 109486881A
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kasugarnycin
fermentation
powder
filter residue
value
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CN109486881B (en
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王亮
祁晨娟
李昶志
潘忠成
李蒲民
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Shaanxi Microbe Bio-Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/44—Preparation of O-glycosides, e.g. glucosides
    • C12P19/46—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin

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  • Organic Chemistry (AREA)
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  • General Health & Medical Sciences (AREA)
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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
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Abstract

The present invention relates to a kind of fermentation medium of kasugarnycin and zymotechnique, the fermentation medium by mass percentage, is made of formula as below: soyabean expeller powder 2.0~5.0%；NaCl:0.1~0.3%；Dried Corn Steep Liquor Powder: 0.2~0.5%；Maltose: 0.8~1.5%；Fish oil: 0.5~1.0%；KH2PO4: 0.03~0.05%；PH value 6.0~7.0 consolidate object content 9~10 kasugarnycin fermentation filter residue: 50~60%；GPE:0.05~0.08%；PH:6.8~7.2.It is effective to realize using kasugarnycin fermentation filter residue by the control of the culture medium and zymotechnique, kasugarnycin fermentation unit is improved, fermentation period shortens, to reach reduction production cost.

Description

A kind of fermentation medium and zymotechnique of kasugarnycin
Technical field
The invention belongs to fermentation technical fields, and in particular to a kind of fermentation medium and zymotechnique of kasugarnycin.
Background technique
Kasugarnycin (Kasugamycin) belongs to produced by springtime streptomycete (Streptomyces kasugaensis) Aminoglycoside antibiotics finds that it is also known as kasugarnycins from Nara County, Japan springtime sacrificial place in April, 1964 within the border.China It was also separated to one plant of kasugarnycin producing strains from the Jiangxi good area of soil in 1963, with Japanese strain in utilization of carbon source and various trainings Supporting cultural characteristic on base has many differences, therefore is named as streptomyces microaureus (S. microaureus).
Kasugarnycin has just been widely used in agricultural since research and development, in Asia and South America.It is to rice blast The prevention and treatment of (including leaf pest, rice head pest, paddy pest) etc. is especially pronounced, generally can reach 80% or more.Moreover, kasugarnycin Applicable crops further include potato, cucumber, celery, sorghum, capsicum, Kidney bean, citrus etc., can prevent and treat the raw tail of beet on beet Carrot soft gull Wen bacterium on spore, potato, the Kidney bean pseudomonas that dwells on Kidney bean, the pseudomonas that sheds tears on cucumber, kind Eggplant leaf mold, cucumber bacterial angular leaf spot etc..
It is noted that meeting modern environmental requirement, by because kasugarnycin is nontoxic to people and animals, noresidue, pollution-free State's green foods development center is recommended as AA grades of pollution-free food production data, is classified as Agricultural product without pollution by the Ministry of Agriculture and recommends Pesticide is classified as Vegetable Standardization engineering by Shanghai City and first elects fungicide.With the raising that people realize chemistry security, spring thunder is mould Plain efficient, wide spectrum, non-harmful biological nature illustrate more and more extensive market prospects.Since it has significant anti-plant Disease fungus activity, therefore be successfully just agriculturally widely used from formulating.Its low toxicity, it is mesh that safety is efficiently, environmentally friendly Has pesticide with broad prospects for development in one of the environment friendly agricultural of preceding most promotion prospect, and current crop pest control One of.
Currently, the fermenting and producing of domestic kasugarnycin uses three grade fermemtation mode, main problem existing for which have with Lower several points:
1, the kasugarnycin fermentation sticky high organic content of filter residue is more intractable, and certain harm is caused to environment.
2, kasugarnycin fermenting and producing higher cost, product lack the market competitiveness.
Summary of the invention
In order to recycle kasugarnycin fermentation filter residue, the production cost of kasugarnycin is reduced, kasugarnycin is effectively improved Fermentation titer, the present invention provides a kind of fermentation medium of kasugarnycin and zymotechniques, pass through the culture medium and fermentation The control of technique, effective to realize using kasugarnycin fermentation filter residue, improve kasugarnycin fermentation unit, fermentation period shortens, To reach reduction production cost.
The present invention adopts the following technical scheme that, a kind of fermentation medium of kasugarnycin, by mass percentage, by following Formula composition: soyabean expeller powder 2.0~5.0%；NaCl:0.1~0.3%；Dried Corn Steep Liquor Powder: 0.2~0.5%；Maltose: 0.8~ 1.5%；Fish oil: 0.5~1.0%；KH2PO4: 0.03~0.05%；PH value 6.0~7.0 consolidates the kasugarnycin fermentation of object content 9~10 Filter residue: 50~60%；GPE:0.05~0.08%；PH:6.8~7.2.
In the preferred embodiments of the present invention, the culture medium the preparation method comprises the following steps: 1. weigh soyabean expeller powder, NaCl, Dried Corn Steep Liquor Powder, maltose, fish oil, KH2PO4, the kasugarnycin fermentation that pH value 6.0~7.0 consolidates object content 9~10 is added Filter residue is uniformly dissolved, and GPE is added, with tap water constant volume to the front volume that disappears；2. being 6.8~7.2 with 30%NaOH tune pH.
A kind of zymotechnique of kasugarnycin, comprising the following steps:
(1) fermentative medium formula: soyabean expeller powder 2.0~5.0%；NaCl:0.1~0.3%；Dried Corn Steep Liquor Powder: 0.2~0.5%； Maltose: 0.8~1.5%；Fish oil: 0.5~1.0%；KH2PO4: 0.03~0.05%；PH value 6.0~7.0 consolidates object content 9~10 Kasugarnycin fermentation filter residue: 50~60%；GPE:0.05~0.08%；PH:6.8~7.2；
(2) soyabean expeller powder, NaCl, Dried Corn Steep Liquor Powder, maltose, fish oil, KH 1. preparation method: are weighed2PO4, pH value 6.0 is added The kasugarnycin fermentation filter residue of~7.0 solid object contents 9~10 is uniformly dissolved, and GPE is added, with tap water constant volume to the front volume that disappears； 2. being 6.8~7.2 with 30%NaOH tune pH；
(3) it sterilizes: being sterilized under 121~125 DEG C of high temperature, sterilize 30min；
(4) be inoculated with: culture medium temperature drops to 30 DEG C, is inoculated with kasugarnycin seed liquor, wherein the inoculum concentration of streptomyces microaureus strain For the volume ratio of 10-20%, fermented and cultured；
(5) ferment: 28~30 DEG C of temperature, ventilatory capacity 1:0.8~1.2,150~200rpm of speed of agitator, tank pressure 0.02~ Ferment 168~170h under the conditions of 0.05MPa, collects fermentation clear liquid purification and obtains tunning；
(6) to tunning filter, take filtrate to get.
In the preferred embodiments of the present invention, pH value 6.0~7.0 consolidates the kasugarnycin fermentation filter of object content 9~10 The mass percent of slag is 50-55%.
In the preferred embodiments of the present invention, in step (4), the inoculum concentration is 10% volume ratio.
In the preferred embodiments of the present invention, fermentative medium formula are as follows: soyabean expeller powder 3.0%；NaCl:0.3%；It is beautiful Rice & peanut milk dry powder: 0.25%；Maltose: 0.8%；Fish oil: 1.0%；KH2PO4: 0.03%；PH value 6.0~7.0 consolidates object content 9~10 Kasugarnycin fermentation filter residue: 55%；GPE:0.08%；PH6.8 is final concentration in the medium above.
The kasugarnycin that the present invention also protects above-mentioned zymotechnique to obtain.
Compared with the culture medium of the prior art and zymotechnique, kasugarnycin fermentation is added in the present invention in the fermentation medium Filter residue, and reduce the dosage of carbon and nitrogen sources, and be utilized kasugarnycin fermentation filter residue to reduce Solid state fermentation pressure.
Using fermentative medium formula provided in the present invention and fermentating controling process, it is mould spring thunder can be greatly improved Plain fermentation unit reduces fermentation costs, and reduces kasugarnycin fermentation filter residue environmental pollution to greatest extent, guarantees it It recycles, realizes that kasugarnycin is stable, efficiently produces.
Zymotechnique of the invention can be such that kasugarnycin fermentation filter residue is recycled, and utilization rate can achieve 80% More than, in general, the mean titre of kasugarnycin is not less than 8000ug/ml in fermentation liquid.
Specific embodiment
The present invention is described in further details below with reference to embodiment, but the present invention is not limited only to following implementations Example.
Embodiment 1
Present embodiment discloses a kind of specific embodiments of fermentation kasugarnycin, include the following steps:
(1) culture medium is prepared, according to kasugarnycin liquid fermentation medium formulation ratio, prepares 1000L culture medium.
Soyabean expeller powder 3.0%；NaCl 0.3% ；Dried Corn Steep Liquor Powder 0.25%；Maltose 0.8%；Fish oil 1.0%； KH2PO4 0.03% ；GPE 0.08%；
PH value 6.20 consolidates kasugarnycin fermentation 50% pH6.80 of filter residue of object content 9.5
(2) culture medium is sterilized 30min under 121 DEG C of high temperature.
(3) culture medium temperature drops to 30 DEG C of inoculation kasugarnycin seed liquors, inoculum concentration 10vol%, fermented and cultured.
(4) 28.0 DEG C of fermentation temperature, ventilatory capacity 1:1, speed of agitator 160rpm, tank press 0.02MPa, under the conditions of ferment 170h collects fermentation clear liquid purification and obtains kasugarnycin.
(5) HPLC method measures the kasugarnycin potency 8976ug/ml of the filtrate.
Embodiment 2
The present embodiment and being different only in that for embodiment 1 have adjusted fermentating formula proportion.
(1) culture medium is prepared, according to kasugarnycin liquid fermentation medium formulation ratio, prepares 1000L culture medium.
Soyabean expeller powder 3.0%；NaCl 0.3%；Dried Corn Steep Liquor Powder 0.25%；Maltose 0.8%；Fish oil 1.0%；KH2PO4 0.03% ；GPE 0.08%；
PH value 6.20 consolidates kasugarnycin fermentation 60% pH6.80 of filter residue of object content 9.5
(2) culture medium is sterilized 30min under 121 DEG C of high temperature.
(3) culture medium temperature drops to 30 DEG C of inoculation kasugarnycin seed liquors, inoculum concentration 10vol%, fermented and cultured.
(4) 28.0 DEG C of fermentation temperature, ventilatory capacity 1:1, speed of agitator 160rpm, tank press 0.02MPa, under the conditions of ferment 170h collects fermentation clear liquid purification and obtains kasugarnycin.
(5) HPLC method measures the kasugarnycin potency 8020ug/ml of the filtrate.
Embodiment 3
The present embodiment and being different only in that for embodiment 1 have adjusted fermentating formula proportion.
(1) culture medium is prepared, according to kasugarnycin liquid fermentation medium formulation ratio, prepares 1000L culture medium.
Soyabean expeller powder 3.0%；NaCl 0.3% ；Dried Corn Steep Liquor Powder 0.25%；Maltose 0.8%；Fish oil 1.0%；KH2PO4 0.03% ；GPE 0.08%
PH value 6.20 consolidates kasugarnycin fermentation 55% pH6.80 of filter residue of object content 9.5
(2) culture medium is sterilized 30min under 121 DEG C of high temperature.
(3) culture medium temperature drops to 30 DEG C of inoculation kasugarnycin seed liquors, inoculum concentration 10vol%, fermented and cultured.
(4) 28.0 DEG C of fermentation temperature, ventilatory capacity 1:1, speed of agitator 160rpm, tank press 0.02MPa, under the conditions of ferment 170h collects fermentation clear liquid purification and obtains kasugarnycin.
(5) HPLC method measures the kasugarnycin potency 9631ug/ml of the filtrate.
Embodiment 4
The present embodiment and being different only in that for embodiment 1 do not add the kasugarnycin fermentation filter that pH value 6.20 consolidates object content 9.5 Slag.
(1) culture medium is prepared, according to kasugarnycin liquid fermentation medium formulation ratio, prepares 1000L culture medium.
Soyabean expeller powder 3.0%；NaCl 0.3%；Dried Corn Steep Liquor Powder 0.25%；Maltose 0.8%；Fish oil 1.0%；KH2PO4 0.03%；GPE 0.08% pH6.80
(2) culture medium is sterilized 30min under 121 DEG C of high temperature.
(3) culture medium temperature drops to 30 DEG C of inoculation kasugarnycin seed liquors, inoculum concentration 10vol%, fermented and cultured.
(4) 28.0 DEG C of fermentation temperature, ventilatory capacity 1:1, speed of agitator 160rpm, tank press 0.02MPa, under the conditions of ferment 170h collects fermentation clear liquid purification and obtains kasugarnycin.
(5) HPLC method measures the kasugarnycin potency 6407ug/ml of the filtrate.
Although above having used general explanation, specific embodiment and test, the present invention is made to retouch in detail It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art 's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed Range.

Claims (7)

1. a kind of fermentation medium of kasugarnycin, which is characterized in that by mass percentage, be made of formula as below: soya bean Dregs of rice powder 2.0~5.0%；NaCl:0.1~0.3%；Dried Corn Steep Liquor Powder: 0.2~0.5%；Maltose: 0.8~1.5%；Fish oil: 0.5~ 1.0%；KH2PO4: 0.03~0.05%；PH value 6.0~7.0 consolidate object content 9~10 kasugarnycin fermentation filter residue: 50~60%； GPE:0.05~0.08%；PH:6.8~7.2.
2. fermentation medium according to claim 1, which is characterized in that the culture medium the preparation method comprises the following steps: 1. claiming Take soyabean expeller powder, NaCl, Dried Corn Steep Liquor Powder, maltose, fish oil, KH2PO4, pH value 6.0~7.0 is added and consolidates object content 9~10 Kasugarnycin fermentation filter residue is uniformly dissolved, and GPE is added, with tap water constant volume to the front volume that disappears；2. being 6.8 with 30%NaOH tune pH ~7.2.
3. a kind of zymotechnique of kasugarnycin, which comprises the following steps:
Fermentative medium formula: soyabean expeller powder 2.0~5.0%；NaCl:0.1~0.3%；Dried Corn Steep Liquor Powder: 0.2~0.5%；Malt Sugar: 0.8~1.5%；Fish oil: 0.5~1.0%；KH2PO4: 0.03~0.05%；PH value 6.0~7.0 consolidates the spring thunder of object content 9~10 Mycin fermentation filter residue: 50~60%；GPE:0.05~0.08%；PH:6.8~7.2；
Preparation method: soyabean expeller powder, NaCl, Dried Corn Steep Liquor Powder, maltose, fish oil, KH are 1. weighed2PO4, addition pH value 6.0~ The kasugarnycin fermentation filter residue of 7.0 solid object contents 9~10 is uniformly dissolved, and GPE is added, with tap water constant volume to the front volume that disappears；② It is 6.8~7.2 with 30%NaOH tune pH；
Sterilizing: sterilizing under 121~125 DEG C of high temperature, and sterilize 30min；
Inoculation: culture medium temperature drops to 30 DEG C, is inoculated with kasugarnycin seed liquor, wherein the inoculum concentration of streptomyces microaureus strain is The volume ratio of 10-20%, fermented and cultured；
Fermentation: 28~30 DEG C of temperature, ventilatory capacity 1:0.8~1.2,150~200rpm of speed of agitator, tank press 0.02~0.05MPa, Under the conditions of ferment 168~170h, collect fermentation clear liquid purification obtain tunning；
(6) to tunning filter, take filtrate to get.
4. zymotechnique according to claim 3, which is characterized in that pH value 6.0~7.0 consolidates the spring thunder of object content 9~10 The mass percent of mycin fermentation filter residue is 50-55%.
5. zymotechnique according to claim 3, which is characterized in that in step (4), the inoculum concentration is 10% volume Than.
6. zymotechnique according to claim 3, which is characterized in that fermentative medium formula are as follows: soyabean expeller powder 3.0%； NaCl:0.3%；Dried Corn Steep Liquor Powder: 0.25%；Maltose: 0.8%；Fish oil: 1.0%；KH2PO4: 0.03%；PH value 6.0~7.0 consolidates object The kasugarnycin fermentation filter residue of content 9~10: 55%；GPE:0.08%；PH6.8 is final concentration in the medium above.
7. the kasugarnycin that the zymotechnique according to any one of claim 3-6 obtains.
CN201811608846.8A 2018-12-27 2018-12-27 Fermentation medium and fermentation process of kasugamycin Active CN109486881B (en)

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Cited By (5)

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Publication number Priority date Publication date Assignee Title
CN111184030A (en) * 2020-01-09 2020-05-22 枣庄市杰诺生物酶有限公司 Kasugamycin raw powder and preparation method and application thereof
CN112210578A (en) * 2020-09-25 2021-01-12 广州市凯卫莎环保科技有限公司 Method for preparing kasugamycin from kitchen waste product
CN113846136A (en) * 2021-11-16 2021-12-28 陕西麦可罗生物科技有限公司 Kasugamycin fermentation medium and fermentation method
CN113862147A (en) * 2021-11-16 2021-12-31 陕西麦可罗生物科技有限公司 Electromagnetic coupling embedded double-tank device and application thereof
CN114540211A (en) * 2020-11-25 2022-05-27 武汉科诺生物科技股份有限公司 Kasugamycin fermentation method

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CN106011201A (en) * 2016-07-01 2016-10-12 宁夏泰瑞制药股份有限公司 Culture medium for producing kasugamycin by fermenting actinomyces microaureus and culture method
CN107815426A (en) * 2016-09-12 2018-03-20 牡丹江佰佳信生物科技有限公司 The special strain therefore of fermenting and producing kasugarnycin and its application
CN107828702A (en) * 2017-12-15 2018-03-23 陕西麦可罗生物科技有限公司 A kind of kasugarnycin fermentation medium and fermentation process

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WO2014165253A1 (en) * 2013-03-12 2014-10-09 Nbip, Llc Compositions and methods for preventing infection of a wound and for advancing the healing process
CN105624240A (en) * 2014-11-28 2016-06-01 牡丹江佰佳信生物科技有限公司 Kasugamycin fermentation medium and fermentation method thereof
CN106011201A (en) * 2016-07-01 2016-10-12 宁夏泰瑞制药股份有限公司 Culture medium for producing kasugamycin by fermenting actinomyces microaureus and culture method
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CN111184030A (en) * 2020-01-09 2020-05-22 枣庄市杰诺生物酶有限公司 Kasugamycin raw powder and preparation method and application thereof
CN112210578A (en) * 2020-09-25 2021-01-12 广州市凯卫莎环保科技有限公司 Method for preparing kasugamycin from kitchen waste product
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CN113846136A (en) * 2021-11-16 2021-12-28 陕西麦可罗生物科技有限公司 Kasugamycin fermentation medium and fermentation method
CN113862147A (en) * 2021-11-16 2021-12-31 陕西麦可罗生物科技有限公司 Electromagnetic coupling embedded double-tank device and application thereof
CN113846136B (en) * 2021-11-16 2023-06-20 陕西麦可罗生物科技有限公司 Kasugamycin fermentation medium and fermentation method
CN113862147B (en) * 2021-11-16 2024-02-27 陕西麦可罗生物科技有限公司 Electromagnetic coupling embedded double-tank device and application thereof

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